RNA Transport Within the Nucleus

细胞核内的 RNA 运输

• 批准号: 6650889
• 负责人: THORU PEDERSON
• 金额: $ 30.21万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6650889
• 关键词: RNA binding protein; cell nucleus; fluorescence microscopy; fluorescent dye /probe; globin; guanosinetriphosphatases; in situ hybridization; intracellular transport; messenger RNA; microinjections; nucleic acid quantitation /detection; oligonucleotides; protooncogene; ribosomal RNA; tissue /cell culture; transcription factor

DESCRIPTION (provided by applicant): This research project involves a team of two cell biologists and a biophysicist who have productively collaborated recently and now are joining together to undertake studies of RNA localization and traffic in the cell nucleus. The proposed research is virtually unique in that intranuclear RNA movements are tracked in living cells, rather than by localization methods that require chemical fixation. The importance of this approach cannot be overstated, since it opens the door to the in vivo investigation of dynamic pathways of intranuclear RNA transport. The two major experimental strategies to be taken, both developed in the P.I?s laboratory, are the fluorescent tagging of endogenous nuclear RNAs and the microinjection of fluorescent RNAs into mammalian cell nuclei. A diffusion-based model for nuclear RNA transport has emerged from this initial work and this will be further investigated. The proposed studies will characterize in detail the intranuclear trafficking patterns of ribosomal RNA as well as beta-globin and c-fos mRNAs. Mutant beta-globin RNAs will be investigated to relate processing defects to intranuclear transport and localization. Studies will also be undertaken of the spliced and polyadenylated but nucleus-restricted RNA, Xist, which is an unusual transcript produced by the inactive X chromosome and is predicted to display very different intranuclear transport properties than mRNAs. Another aspect of the project will address the intranuclear traffic of these RNAs in relation to proteins known to be involved in nuclear import or RNA export, i.e. the Ran-GTPase and TAP.

描述（由申请人提供）：该研究项目涉及一个团队 两位细胞生物学家和一位生物物理学家进行了卓有成效的合作 最近和现在正在联合进行RNA定位的研究 和细胞核中的交通。拟议的研究实际上是独一无二的 追踪活细胞中的核内 RNA 运动，而不是通过需要化学固定的定位方法。这一点的重要性 这种方法怎么强调都不为过，因为它打开了体内研究的大门 核内RNA转运动态途径的研究。两大主要 要采取的实验策略，都是在 P.I 实验室开发的， 是内源性核RNA的荧光标记和显微注射 荧光RNA进入哺乳动物细胞核。基于扩散的模型 核 RNA 运输已经从这项初步工作中出现，这将是 进一步调查。拟议的研究将详细描述 核糖体 RNA 以及 β-珠蛋白的核内运输模式 c-fos mRNA。将研究突变 β-珠蛋白 RNA 的相关加工 核内运输和定位缺陷。研究也将 进行剪接和聚腺苷酸化但核限制性 RNA，Xist， 这是由不活跃的 X 染色体产生的不寻常的转录本， 预计显示出与 mRNA。该项目的另一个方面将解决核内交通问题 这些 RNA 与已知参与核输入的蛋白质有关或 RNA 输出，即 Ran-GTPase 和 TAP。